马铃薯卷叶病毒外壳蛋白基因克隆转化 及其转基因后代的表达

南相日,刘文萍,刘琦,夏善勇

中国农学通报. 2007, 23(7): 106-106

中国农学通报 ›› 2007, Vol. 23 ›› Issue (7) : 106-106. DOI: 10.11924/j.issn.1000-6850.0707106
生物技术科学

马铃薯卷叶病毒外壳蛋白基因克隆转化 及其转基因后代的表达

  • 南相日,刘文萍,刘琦,夏善勇
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Cloning and Transformation and Expression of Potato leafroll virus coat protein gene in Potato

  • Nan Xiangri, Liu Wenping, Liu Qi, Xia Shanyong
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摘要

根据GenBank中的马铃薯卷叶病外壳蛋白基因(PLRV-CP)全序列，设计一对特异性引物，以马铃薯卷叶病毒RNA为模板，克隆了马铃薯卷叶病外壳蛋白基因，在pBI121的基础上构建了植物表达载体。利用PLO(Poly-L-Ornithine)将PLRV-CP基因导入到马铃薯加工型品种大西洋的原生质体中，获得了转基因后代。在5株转化后代中扩增出长度为610bp的目标DNA片断，说明导入的外源PLRV-CP基因已经整合到马铃薯基因组中。Southern blot分析结果进一步证明了PCR结果的正确性。RT-PCR结果表明，在3株转化后代叶片中具有阳性表达。马铃薯卷叶病毒接种实验结果表明，转基因植株比对照有明显的马铃薯卷叶病抗性

Abstract

A pair of specific primer had been designed according to full length sequence of Potato leafroll virus coat protein gene (PLRV-CP) in the GenBank. Potato leafroll virus coat protein gene (PLRV-CP) had been cloned with the RNA of Potato leafroll virus as moplate, and plant expression vector had been constructed. PLRV-CP gene had been transformed to protoplate of the potato variety Daxiyang by the method of PLO (Poly-L-Ornithine) and transgenic plants. About 610bp interest DNA fragment had been amplification in 5 transgenic plants. This showed that exogenous PLRV-CP had been integrated to the genomy of potato. And the result of the Southern blot proved the correct of the PCR and the result of the RT-PCR further demonsticated that: there are positive expression in the 3 progenies of transgenic plants. And the potato leafroll virus inoculation experiment show that transgenic plants Resistant to the potato leafroll compared with control plants.

关键词

马铃薯；PLRV-CP基因；原生质体；克隆

Key words

Potato;PLRV-CP gene;Protoplast;Clone

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南相日,刘文萍,刘琦,夏善勇. 马铃薯卷叶病毒外壳蛋白基因克隆转化 及其转基因后代的表达. 中国农学通报. 2007, 23(7): 106-106 https://doi.org/10.11924/j.issn.1000-6850.0707106
Nan Xiangri, Liu Wenping, Liu Qi, Xia Shanyong. Cloning and Transformation and Expression of Potato leafroll virus coat protein gene in Potato. Chinese Agricultural Science Bulletin. 2007, 23(7): 106-106 https://doi.org/10.11924/j.issn.1000-6850.0707106

参考文献

[1]Robert R, Martin Paul K, Keese Mark J, et al. Evolution and molecular biology of luteoviruses. Annu Rev Phytopathol, 1990,28:341-363.
[2]张荣信,哈斯阿古拉,张鹤龄.马铃薯卷叶病毒56kD蛋白基因及3’端非编码区克隆和序列分析.病毒学报,1997,13(3):247-254.
[3]Barker H, Reavy B, Webster K D, et al. Relationship between transcript production and virus resistance in transgenic tobacco expressing the potato leafroll virus coat protein gene. Plant Cell Reports, 1993,13:54-58.
[4]张鹤龄,李天然,哈斯阿占拉,等.抗卷叶病毒(PLRV)转基因马铃薯栽培种及其抗病性的研究.病毒学
报,1995,13:54-58.
[5]赵福宽,张鹤龄,李天然,等.表达PLRV CP基因的转基因马铃薯抗病性田间鉴定.北京农学院学报,1997,12(3):17-19.
[6]Gould A R. Studies on encapsidated viroid-like RNA, II. Purification and characterization of a viroid-like RNA associated with velvet tobacco mottle virus (VT-MOV ).virology, 1981,108:123-133.
[7]J.萨姆布鲁克,E.F.费里奇,T.曼尼阿蒂斯著分子克隆实验指南.北京:科学出版社,1992:19-22.
[8]卫志明,许智宏.大豆原生质体培养和植株再生.植物学报,1990,32(8):582-588.
[9]南相日,卫志明.利用聚鸟氨酸提高大豆原生质体外源基因的转化效率.实验生物学报,1999,32(4):409-413.
[10]Stewart CN Jr. Via LE A rapid CTAB DNA isolation technique useful for RAPD fingerprinting and other PCR applications. Biotechniques, 1993,14:748-751.

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